• Title/Summary/Keyword: Soil DNA

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Construction and Characterization of a cDNA Library from the Camelina sativa L. as an Alternative Oil-Seed Crop (신 바이오디젤 원료 작물인 Camelina의 cDNA library 제작 및 유전자 특성)

  • Park, Won;Jang, Young-Seok;Ahn, Sung-Ju
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.55 no.2
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    • pp.151-158
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    • 2010
  • Camelina sativa L., known as popular names "gold-of-pleasure" or "false flax" is an alternative oilseed crop that can be grown under different climatic and soil conditions. Up to date, however, the genomic information of Camelina has not been studied in detail. Therefore, a cDNA library was constructed and characterized from young leaves. The constructed cDNA library incorporated of 1334 cDNA clones and the size of the insertion fragments average was 736 base pair. We generated a total of 1269 high-quality expressed sequence tags (ESTs) sequences. The result of cluster analysis of EST sequences showed that the number of unigene was 851. According to subsequent analysis, the 476 (55.9%) unigenes were highly homologous to known function genes and the other 375 (44.1%) unigenes were unknown. Remaining 63 (7.4%) unigenes had no homology with any other peptide in NCBI database, indicating that these seemed to be novel genes expressed in leaves of Camelina. The database-matched ESTs were further classified into 17 categories according to their functional annotation. The most abundant of categories were "protein with binding function or cofactor requirement (27%)", "metabolism (11%)", "subcellular localization (11%)", "cellular transport, transport facilities and transport routes (7%)", "energy (6%)", "regulation of metabolism and protein function (6%)". Our result in this study provides an overview of mRNA expression profile and a basal genetic information of Camelina as an oilseed crop.

A Thermostable Protease Produced from Bacillus sp. JE 375 Isolated from Korean Soil (한국의 토양으로부터 내열성 단백질 분해효소를 생산하는 Bacillus sp. JE 375의 선별)

  • Kim, Ji-Eun;Bai, Dong-Hoon
    • Korean Journal of Food Science and Technology
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    • v.38 no.3
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    • pp.419-426
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    • 2006
  • A thermophilic microorganism, strain JE 375, which produces a thermostable protease, was isolated from soil and compost in Korea. This gram-positive, rod-shaped, catalase positive, motility positive, and hemolysis ${\beta}$ containing organism was implicated in glucose fermentation, mannitol fermentation, xylose oxidation, aerobic activity and spore formation. The color of the colony was yellowish white. The temperature range for growth at pH 6.5 was between 55 and $70^{\circ}C$, with an optimum growth temperature of $65^{\circ}C$. This result confirmed the strain JE 375 as a thermophilic microorganism. The enzyme was produced aerobically at $65^{\circ}C$ during 20 hr in a medium (pH 6.5) containing 1% trypton. 1% maltose, 0.5% yeast extract and 1% NaCl. The 16S rDNA of strain JE 375 had 97.6% sequence similarity with the 16S rDNA of Bacillus caldoxyloyticus. On the basis of biochemical and physiological properties and phylogenetic analysis, we named the isolated strain as Bacillus sp. JE 375. The thermostable protease from Bacillus sp. JE 375 had been partially purified and characterized. The molecular weight of the enzyme was deduced from SDS-PAGE and gel chromatography as 55 kDa and its optimal temperature was $60^{\circ}C$. The enzyme showed its highest activity at pH 7.5 and was stable from pH 7.0 to 8.0.

Functional Genomics for Mass Analysis of Useful Genes in Panax ginseng C.A. Meyer (인삼의 유용유전자원 확보를 위한 기능 유전체연구)

  • Yang, Deok-Chun
    • Proceedings of the Ginseng society Conference
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    • 2004.05a
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    • pp.17-28
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    • 2004
  • As Korean ginseng is hybrid, an individual variation is very severe, and it takes long times in new breeding because it is required 4 years to pick the seed. But, transformation technique makes the high-functional breeding in short time. The focus of these ginseng studies is to find and secure the useful gene. And it is urgent to accumulate the fundamental data for the molecular breeding and secure the useful genes. Therefore, transformation and soil acclimatization technique are necessary to molecular breeding in use of the introduction of functional genes. In this study, it add to secure of new regulation gene and useful gene as to accumulate the fundamental data for the place where it will contribute to raise the national competitive power. To analyze the useful genes in large scale, we constructed CDNA libraries with various tissues, species, and treated tissue. EST analysis of ginseng perform in large scale and build the EST database of ginseng. We perform the full length sequencing about the selected lots of clones that include the entire open reading frame of the amino acid residues and construct cDNA chip with the parental EST clones. Establishment of the transformation and a soil acclimatization system throuth the re-introduction of the selected ginseng gene that related with the secondary metabolism and anti-stress into the ginseng.

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Sequence-Based Screening for Putative Polyketide Synthase Gene-Harboring Clones from a Soil Metagenome Library

  • JI SANG CHUN;KIM DOCKYU;YOON JUNG-HOON;OH TAE-KWANG;LEE CHOONG-HWAN
    • Journal of Microbiology and Biotechnology
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    • v.16 no.1
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    • pp.153-157
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    • 2006
  • A soil metagenomic library was constructed using an E. coli-fosmid cloning system with environmental DNAs extracted from Kwangreung forest topsoil. We targeted the genes involved in the biosynthesis of bacterial polyketides. Initially, a total of 36 clone pools (10,800 clones) were explored by the PCR-based method using the metagenomic DNAs from each pool and a degenerate primer set, which has been designed based on the highly conserved regions among ketoacyl synthase (KS) domains in actinomycete type I polyketide synthases (PKS Is). Six clone pools were tentatively selected as positive and further examined through a hybridization-based method for selecting a fosmid clone containing PKS I genes. Colony hybridization was performed against fosmid clones from the 6 positive pools, and finally 4 clones were picked out and confirmed to contain the conserved DNA fragment of KS domains. In this study, we present a simple and feasible sorting method for a desired clone from metagenomic libraries.

Cohnella panacarvi sp. nov., a Xylanolytic Bacterium Isolated from Ginseng Cultivating Soil

  • Yoon, Min-Ho;Ten, Leonid N.;Im, Wan-Taek
    • Journal of Microbiology and Biotechnology
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    • v.17 no.6
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    • pp.913-918
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    • 2007
  • A Gram-positive, aerobic, rod-shaped, nonmotile, endospore-forming bacterium, designated Gsoil $349^T$, was isolated from soil of a ginseng field and characterized using a polyphasic approach. Comparative analysis of 16S rRNA gene sequences revealed that the strain Gsoil $349^T$ belongs to the family Paenibacillaceae, and the sequence showed closest similarity with Cohnella thermotolerans DSM $17683^T$ (94.1%) and Cohnella hongkongensis DSM $17642^T$ (93.6%). The strain showed less than 91.3% 16S rRNA gene sequence similarity with Paenibacillus species. In addition, the presence of MK-7 as the major menaquinone and $anteiso-C_{15:0},\;iso-C_{16:0},\;and\;C_{16:0}$ as major fatty acids suggested its affiliation to the genus Cohnella. The G+C content of the genomic DNA was 53.4 mol%. On the basis of its phenotypic characteristics and phylogenetic distinctiveness, strain Gsoil $349^T$ should be treated as a novel species within the genus Cohnella for which the name Cohnella panacarvi sp. nov. is proposed. The type strain is Gsoil $349^T\;(=KCTC\;13060^T=\;DSM\;18696^T)$.

Use of Terminal Restriction Length Polymorphism (T-RFLP) Analysis to Evaluate Uncultivable Microbial Community Structure of Soil

  • Chauhan, Puneet Singh;Shagol, Charlotte C.;Yim, Woo-Jong;Tipayno, Sherlyn C.;Kim, Chang-Gi;Sa, Tong-Min
    • Korean Journal of Soil Science and Fertilizer
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    • v.44 no.1
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    • pp.127-145
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    • 2011
  • Various environmental ecosystems are valuable sources for microbial ecology studies, and their analyses using recently developed molecular ecological approaches have drawn significant attention within the scientific community. Changes in the microbial community structures due to various anthropogenic activities can be evaluated by various culture-independent methods e.g. ARISA, DGGE, SSCP, T-RFLP, clone library, pyrosequencing, etc. Direct amplification of total community DNA and amplification of most conserved region (16S rRNA) are common initial steps, followed by either fingerprinting or sequencing analysis. Fingerprinting methods are relatively quicker than sequencing analysis in evaluating the changes in the microbial community. Being an efficient, sensitive and time- and cost effective method, T-RFLP is regularly used by many researchers to access the microbial diversity. Among various fingerprinting methods T-RFLP became an important tool in studying the microbial community structure because of its sensitivity and reproducibility. In this present review, we will discuss the important developments in T-RFLP methodology to distinguish the total microbial diversity and community composition in the various ecosystems.

Effects of habitat differences on the genetic diversity of Persicaria thunbergii

  • Nam, Bo Eun;Nam, Jong Min;Kim, Jae Geun
    • Journal of Ecology and Environment
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    • v.40 no.2
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    • pp.84-88
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    • 2016
  • To understand the effects of habitat characteristics on the genetic diversity of Persicaria thunbergii, three sites of different environmental conditions in a water system were surveyed. Site A was the closest to the source of the water system, and there was a dam between sites A and B. Site C is located on the lowest downstream in the water system. Vegetation survey of four quadrats at each site was performed, and soil samples were collected for physicochemical analysis. Random amplification of polymorphic DNA (RAPD) analysis of ten P. thunbergii individuals at each site was conducted to calculate population genetic diversity and genetic distance among populations. Soil was sterile sand at site A, whereas loamy soil at sites B and C. A pure stand of P. thunbergii appeared at site A, while other species occurred together (such as Humulus japonicus and Phragmites australis) at sites B (Shannon-Wiener index; $H_B=0.309$) and C ($H_C=0.299$). Similar to the species diversity, genetic diversity (Nei's gene diversity; h) within population of site A ($h_A=0.2381$) was relatively lower than sites B ($h_B=0.2761$) and C ($h_C=0.2618$). However, site C was separated from sites A and B in genetic distance rather than the geographical distance (Nei's genetic distance; A~B, 0.0338; B~C, 0.0685; A~C, 0.0833).

Screening and Identification of Antibacterial Actinomycetes against Bacteria Causing Rusty Root on Ginseng (인삼의 적변을 유발하는 세균에 대하여 항균활성을 가지는 방선균 선발 및 동정)

  • Han, Sung-Hee;Ryu, Dong-Kul;Choi, Seung-Hyun;Choi, Jae-Eul;An, Gilh-Wan
    • Korean Journal of Agricultural Science
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    • v.37 no.2
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    • pp.255-260
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    • 2010
  • Rusty root, the browning disease on ginseng, decreases quality and value. Recent studies indicated that endophytic bacteria could be a possible cause of rusty root. Actinomycetes antagonistic to the rusty-root-causing bacteria were isolated from soil. Twenty nine out of 932-isolates of Actinomycetes from soil showed antibacterial activity against Agrobacterium tumefaciens and Pseudomonas veronii an endophytic isolate in ginseng. The strongest antibacterial strain(ATO4O104) was classified based on 16S rDNA sequence. The Actinomycetes strain, ATO4O104, isolated in soil of USA volcano national park was identified as Streptomyces adephospholyticus. To test plant toxicity, radish seeds were sprouted with the culture of S. adephospholyticus and it did not show any harmful effect. The butanol partition out of n-hexane, ethyl acetate, butanol, and water partions showed the highest antibacterial activity.

Mapping of Quantitative Trait Loci for Yield and Grade Related Traits in Peanut (Arachis hypogaea L.) Using High-Resolution SNP Markers

  • Liang, Yuya;Baring, Michael R.;Septiningsih, Endang M.
    • Plant Breeding and Biotechnology
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    • v.6 no.4
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    • pp.454-462
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    • 2018
  • Yield and grade are the key factors that affect production value of peanut. The objective of this study was to identify QTLs for pod yield, hundred-seed weight, and total sound mature kernel (TSMK). A total of 90 recombinant inbred lines, derived from Tamrun OL07 and a breeding line Tx964117, were used as a mapping population and planted in Brownfield and Stephenville, Texas. A genetic map was developed using 1,211 SNP markers based on double digest restriction-site associated DNA sequencing (ddRAD-seq). A total of 10 QTLs were identified above the permutation threshold, three for yield, three for hundred-seed weight and four for TSMK, with LOD score values of 3.7 - 6.9 and phenotypic variance explained of 12.2% - 35.9%. Among those, there were several QTLs that were detected in more than one field experiment. The commonly detected QTLs in this study may be used as potential targets for future breeding program to incorporate yield and grade related traits through molecular breeding.

First Report of Allantophomopsiella pseudotsugae Isolated from Soil in Korea

  • Wajihi, Ally Hassan;Lee, Seung-Yeol;Das, Kallol;Eom, Ahn-Heum;Jung, Hee-Young
    • The Korean Journal of Mycology
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    • v.47 no.1
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    • pp.29-34
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    • 2019
  • A fungal isolate designated 17E029 was isolated from a soil sample in Jeju, Korea. The strain was similar to other Allantophomopsiella species in its morphological characteristics such as grey mycelia, conidiophore, and conidia sizes. The isolate produced aerial mycelia, which appeared grey on the reverse side of the media surfaces and turned black on the front side of the colonies. The conidiophores emanating from the hyphae were hyaline, grey, aseptate, branched, and $6.7{\sim}9.2{\times}1.8{\sim}2.5{\mu}m$. Conidiogenous cells were ovoid to subcylindrical, discrete, guttulate, and hyaline. Conidia were hyaline, aseptate, smooth, guttulate, oval to subcylindrical, irregular in shape, and $6.0{\sim}7.8{\times}3.0{\sim}3.4{\mu}m$. The strain was confirmed based on phylogenetic analysis of the closest related organism, A. pseudotsugae CBS 288.37, using the partial 28S, internal transcribed spacer rDNA regions, and partial RNA polymerase II second largest subunit locus (RPB2) gene sequences along with its culture characteristics. Therefore, morphological observations and phylogenetic analysis revealed that strain 17E029 is similar to the previously identified A. pseudotsugae. Hence, this species was described as A. pseudotsugae strain 17E029, which is a new record in Korea.